638 resultados para Keith Muchowski


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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Background: Centromeres are essential for chromosome segregation, yet their DNA sequences evolve rapidly. In most animals and plants that have been studied, centromeres contain megabase-scale arrays of tandem repeats. Despite their importance, very little is known about the degree to which centromere tandem repeats share common properties between different species across different phyla. We used bioinformatic methods to identify high-copy tandem repeats from 282 species using publicly available genomic sequence and our own data.Results: Our methods are compatible with all current sequencing technologies. Long Pacific Biosciences sequence reads allowed us to find tandem repeat monomers up to 1,419 bp. We assumed that the most abundant tandem repeat is the centromere DNA, which was true for most species whose centromeres have been previously characterized, suggesting this is a general property of genomes. High-copy centromere tandem repeats were found in almost all animal and plant genomes, but repeat monomers were highly variable in sequence composition and length. Furthermore, phylogenetic analysis of sequence homology showed little evidence of sequence conservation beyond approximately 50 million years of divergence. We find that despite an overall lack of sequence conservation, centromere tandem repeats from diverse species showed similar modes of evolution.Conclusions: While centromere position in most eukaryotes is epigenetically determined, our results indicate that tandem repeats are highly prevalent at centromeres of both animal and plant genomes. This suggests a functional role for such repeats, perhaps in promoting concerted evolution of centromere DNA across chromosomes.

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Objective: Local invasion of bone is a frequent complication of oral squamous cell carcinoma (OSCC). Development of these osteolytic lesions is mediated by osteoclasts. Receptor activation of NF-kappa B ligand (RANKL) signaling, counteracted by osteoprotegerin (OPG), regulates osteoclastogenesis. Previous studies in rodent models have demonstrated that inhibition of RANKL decreases tumor growth and lesions within bone. However, the contributory role of OSCC cells to this disease process has yet to be defined.Methods: RANKL expression was assessed in a panel of OSCC cell lines by qPCR, flow cytometry, and ELISA. Induction of osteoclastogenesis was assessed by co-culture with macrophages or with OSCC-derived conditioned medium. In an animal model of bone invasion, nude mice were injected intratibially with UMSCC-11B cells expressing a RANKL luciferase promoter to detect tumor-derived RANKL activity. Osteolytic lesions were analyzed by X-ray, micro-CT, and histological methods. RANKL expression was assessed in human OSCC tissues by immunohistochemistry.Results: We demonstrated that OSCCs express varied levels of all RANKL isoforms, both membrane-bound and soluble RANKL. Both co-culture and treatment with OSCC-conditioned media induced osteoclastogenesis. In mice, we demonstrated human RANKL promoter activity during bone invasion. Over the course of the experiment, animals suffered osteolytic lesions as RANKL-driven luciferase expression increased with time. After 8 weeks, human-derived RANKL was detected in areas of bone resorption by immunohistochemistry. Similar epithelial RANKL expression was detected in human OSCC tissues.Conclusion: These data demonstrate the ability of OSCCs to produce RANKL, directly altering the tumor microenvironment to increase osteoclastogenesis and mediate local bone invasion. (C) 2012 Elsevier Ltd. All rights reserved.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Objective. The purpose of this study was to evaluate the presence of myofibroblasts, frequently associated with a more aggressive neoplastic behavior, in oral tongue squamous cell carcinoma (TSCC) of young patients and to compare with the distribution observed in older patients.Study Design. Tumor samples from 29 patients younger than 40 years old affected by TSCC were retrieved and investigated for the presence of stromal myofibroblasts by immunohistochemical reactions against a smooth muscle actin, and the results obtained were compared to TSCC cases affecting older patients.Results. No positive reaction could be found in the stromal areas devoid of neoplastic tissue, whereas myofibroblasts were present in 58.6% of the lesions in young patients and in 75.9% of the older ones. No significant difference was found when comparing the invasive front and the overall stroma of both groups, and no correlation could be obtained with stromal a smooth muscle actin expression, higher tumor grades or clinical stage (P > .05).Conclusion. There was no significant difference between the presence of stromal myofibroblasts of TSCC affecting young and old individuals.

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p38 mitogen-activated protein kinases (MAPKs) are critical for innate immune signaling and subsequent cytokine expression in periodontal inflammation and bone destruction. In fact, previous studies show that systemic p38 MAPK inhibitors block periodontal disease progression. However, development of p38 MAPK inhibitors with favorable toxicological profiles is difficult. Here, we report our findings regarding the contribution of the downstream p38 MAPK substrate, mitogen-activated protein kinase-activated protein kinase 2 (MK2 or MAPKAPK-2), in immune response modulation in an experimental model of pathogen-derived lipopolysaccharide (LPS)-induced periodontal bone loss. To determine whether small interfering RNA (siRNA) technology has intraoral applications, we initially validated MK2 siRNA specificity. Then, gingival tissue surrounding maxillary molars of rats was injected with MK2 siRNA or scrambled siRNA at the palatal regions of bone loss. Intraoral tissues treated with MK2 siRNA had significantly less MK2 mRNA expression compared with scrambled siRNA-treated tissues. MK2 siRNA delivery arrested LPS-induced inflammatory bone loss, decreased inflammatory infiltrate, and decreased osteoclastogenesis. This proof-of-concept study suggests a novel target using an intraoral RNA interference strategy to control periodontal inflammation.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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PURPOSE: Our aim was to determine whether the mucosa of a canine jejunal pouch used as a rectal substitute after proctocolectomy retains its morphologic features better than the mucosa of a canine ileal pouch so used. METHODS: Among ten dogs that underwent proctocolectomy, five had a jejunal pouch-distal rectal anastomosis and five an ileal pouch-distal rectal anastomosis. After six months, the animals were killed, the intestinal pouches and portions of unaltered distal ileum were removed, and a blinded, mucosal morphometric analysis was performed. RESULTS: The mucosa of the jejunal pouches had an overall thickness (mean ± standard deviation, anterior and posterior walls, 1,300 ± 140 μm), villous height (286 ± 46 μm), and crypt depth (790 ± 77 μm) greater than that of the ileal pouches (920 ± 170, 208 ± 47, and 530 ± 130 μm, respectively; P < 0.05). Moreover, the mucosal thickness of the jejunal pouches was similar to that of the distal ileum proximal to the pouch (1,200 ± 200 μm; P > 0.05), whereas the mucosal thickness of the ileal pouch was thinner (P < 0.05). CONCLUSIONS: The jejunal mucosa retains its major morphometric features when the jejunum is used as a rectal substitute after proctocolectomy. In contrast, the ileal mucosa atrophies when the ileum is so used.

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Pós-graduação em Genética e Melhoramento Animal - FCAV

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A survey of 41 mule deer (Odocolleus hemionus) and three white-tailed deer (O. virginianus) for bovine tuberculosis was conducted on a Montana (USA) cattle ranch from 2 November 1993 through January 1994. Gross and microscopic lesions typical of tuberculosis were present in tonsil and lymph nodes of the head, thorax, and abdomen of one adult female mule deer. Additionally, a single microgranuloma considered morphologically suggestive of tuberculosis was present in one lymph node of the head of a second mule deer. Mycobacterial isolates from lymph nodes of the head and thorax of the first deer were identified as Mycobacterium bovis.

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Charles Adams (Faculty Advisor), Delbert Kuhlman, John Klingenberg, Ardyce Haring, Harvey Jorgensen, Roy Volzke, Billie Reed, Paul Yeutter Prof. Loeffel, Carolyn Hall, Byron Kort, Larry Paul, Elwin Mosier, Charles Corkle, Kay Robohm, Daniel Stilwell Duane Stokebrand, Gary Briggs, Walt Patterson, Wendell Mousel, Keith Smith, Darrel Zessin, Richard Bonne, Donald Kasbohm, Bruce Skinner Bob Discoe, Doyle Hulme, Jim Smith, Carl Lorenzen, Jay Cook, Gary Berke, Bob Volk, Roger Hild Donald Kuhl, Russell Person, Ray Cada, Ray DeBower, Bob Dannert, Phil Starck, Kay Knudsen, Jerry Brownfield, Allan McClure, Wally Bierman Morris Ochsner, Warren Mitchell, Ed McReynolds, Gerald Dart, Arza Snyder, Mervy Schliefert, Arley Waldo, Tom Hoffman, John Wink, Virgil Gellermann, Duane Neuman

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President Roger Wehrbein Vice President Ted Klug Secretary George B. O'Neal Treasurer Ralph Hazen Marshal Bud Reece Historian Tom Kraeger Co-Historian John Zauha Ag. Executive Representative Larry Williams Faculty Advisor Dr. E. B. Peo, Jr. George Ahlschwede Richard Hahn Henry Beel Ralph Hazen Gary Briggs Gary Heineman Leslie Cook Max Hauser Richard Eberspacher Buce Jameson Russ Edeal Leon Janovy William Ehresman Alan Jorgensen Rolland Eubanks John Joyner Mickey Evertson Marshall Jurgens Jesse Felker Ron Kahle Mylon Filkins Donald Kavan Richard Frahm Max Keasling Roger French Ronald Kennedy Angus Garey Ted Klug Ed Gates Herb Kraeger Gerald Gogan Tom Kraeger Gerald Goold Fernando Lagos Jay Graf Gerald Lamberson Lloyd Langemeier Ralph Langemeier Gerald Loseke Donald Meiergerd Lowell Minert John Oeltjen George B. O'Neal Don Ormesher Larry Ott Bud Reece Ron Sabatka Keith Smith Ronald Smith Donn Simonson Daryl Starr Galen Stevens Eugene Turdy Ernest Thayer Charles Thompson Jerry Thompson Eli Thomssen William Watkins Allen Trumble Robert Weber Lawrence Turner Dan Wehrbein Reginald Turner Roger Wehrbein Vance Uden Dick White Max Waldo Billy Williams Blair Williams Larry Williams D. Patrick Wright John Zauha

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Stress hormones in Rocky Mountain bighorn sheep (Ovis canadensis canadensis), produced in response to environmental changes, road development, or high population density, may impact their immune systems to a threshold level that predisposes them to periodic, large-scale mortality. We compared the stress response to a novel environmental situation and repeated handling between bighorn sheep born and raised in captivity (CR) and bighorn sheep born in the wild (WC) and brought into captivity. We measured plasma epinephrine, norepinephrine, cortisol, and fecal glucocorticoid metabolites (FGM). Three weeks after each group’s arrival we used a one-time drop-net event to elicit an acute stress response, and we collected blood samples from each sheep over 35 minutes, as well as one fecal sample. We collected blood and fecal samples from both groups on 7 other occasions over the subsequent 6 months. We also collected fecal samples from the pen at approximately 24-hour intervals for 3 days following every handling event to monitor the stress response to handling. We found that CR sheep had a stronger autonomic nervous system response than WC sheep, as measured by epinephrine and norepinephrine levels, but we found a very similar hypothalamic–pituitary–adrenal axis (HPA) response, measured by cortisol levels, to the acute stress event of a drop-net restraint. We also found that once the WC sheep had acclimated, as indicated by the return to the initial baseline FGM levels within 12 weeks, the CR and WC groups’ HPA responses to sampling events were not significantly different from one another. Fecal samples can provide a noninvasive mechanism for managers to monitor baseline FGM for a given herd. Using long-term monitoring of FGM rather than values from a single point in time may allow managers to correlate these levels to outside influences on the herd and better understand the impacts of management changes, population density, or increased human developments on the health of the sheep population.